In order to improve the cracking resistance of reinforced concrete and give full play to the advantages of prefabricated assembly structure in construction, prestressed reinforced concrete composite beam (PRCC) is proposed. Through the bending static test of seven I-shaped beam specimens, the bending failure modes and bearing capacity of PRCC and reinforced concrete composite beam are compared and analyzed, and the effects of prestress size, prestressed reinforcement layout and prestress application sequence on the flexural behavior of PRCC beams are studied. The results show that the cracking load and ultimate load of PRCC beams significantly increased after prestressing, and prestressed tendons can effectively control the crack development. With the increase of prestressing degree, the deformation resistance and bending stiffness of PRCC beams are increased. The application sequence of prestress has little influence on the mechanical properties of PRCC beams. The crack width, stiffness and normal section bearing capacity of PRCC beam are analyzed, and the calculated results are in good agreement with the experimental results.